Figure 3. Essential 8 Flex medium limits fluctuations in levels of active FGF2. Measurements of heat-mediated loss of active FGF2 were used to simulate active FGF2 levels in PSC cultures over the course of a week.
With a better than standard stem cell media
Test the limits of your research and pioneer your own breakthrough using a stem cell media you know you can trust. Supported by a proven track record of results.
Since the breakthrough of generating iPSCs in 2007, stem cell research has exploded!
Creating unrivalled possibilities in the understanding, treatment and potential cure of diseases once thought of as untreatable, research into the medical use of stem cells is revolutionary.
With our mission to make the world safer, cleaner and healthier we have always been at the forefront of harnessing these powerful innovations into a total system covering your stem cell research workflow.
For over a decade, Gibco™ have provided the key tools and resources you need to address challenges in your stem cell research. Our complete portfolio of products and services can support every step of your research and you can benefit from the continuity, efficiency, and precision that drive discovery.
See the links below for more information. Then connect with our expert sales team to see the results first hand.
Protocols
Guidebooks
Scientific Posters
Publications
Application notes
Videos
FAQs
Pluripotent stem cell (PSC) research requires careful attention to culture conditions. Gibco media and reagents have been at the forefront of pluripotent stem cell research for years. Gibco products deliver culture with confidence.
Find the right solution for your stem cell culture needs
| Product | StemFlex Medium | Essential 8 Medium | Essential 8 Flex Medium | |
| Ideal for | Robust maintenance of PSC culture, difficult cell lines, and superior performance in single cell passaging and gene editing | Consistent PSC culture and superior reprogramming | PSC culture with flexible feeding schedule | |
| Defined | Animal origin components (BSA) | Xeno-free; no animal derived components, human derived | Xeno-free; no animal-derived components; human derived | |
| Recommended cell types | Human PSCs | Human PSCs | Human PSCs | |
| Weekend-free feeding schedule | Yes | No | Yes | |
| Genome editing | Best | Fair | Fair | |
| Robustness | Best | Fair | Good | |
| Recommended matrix | Geltrex Matrix rhLaminin-521 | Vitronectin (VTN-N) Recombinant Protein rhLaminin-521 | Vitronectin rhLaminin-521 | |
| Recommended level of dissociation and passaging reagent | Clump | Versene Solution | Versene Solution | Versene Solution |
| Small cluster | StemPro Accutase Cell Dissociation Reagent, RevitaCell Supplement helpful but not required | StemPro Accutase Cell Dissociation Reagent, optional RevitaCell Supplement added to medium during first 18-24 hours post-passage for improved recovery | StemPro Accutase RevitaCell Supplement | |
| Single cell | TrypLE Select or Tryple Express Enzyme, RevitaCell Supplement helpful but not required during recovery if using rhLaminin-521 | TrypLE Select or Tryple Express Enzyme with RevitaCell Supplement added to medium during first 18-24 hours post-passage | TrypLE Select or Tryple Express Enzyme with RevitaCell Supplement | |
Essential 8—Routine PSC expansion and maintenance
Proven to maintain pluripotency in multiple iPSC lines, Essential 8 Medium has been used to scale up production of iPSCs—supporting iPSC growth for >50 passages with no signs of karyotypic abnormalities and maintaining the ability of iPSCs to differentiate into all three germ line lineages.
iPSC Morphology—iPSCs cultured in Essential 8 medium on vitronectin at P4.
Staining—Tra-1-60 staining of iPSCs cultured in Essential 8 medium on vitronectin at P13.
iPSCs grown on Essential 8 Medium maintain pluripotency and retain normal karyotype. CDI iPSC line shown here at p18 on Essential 8 Medium and Vitronectin.
Superior recovery during adaption
rhLaminin-521 has been proven to promote cellular survival under stressful conditions in the absence of small molecule inhibitors (Figure 1). In addition, rhLaminin-521 has been shown to support PSC growth for >20 passages without any signs of karyotypic abnormalities and to maintain the ability of PSCs to differentiate into all three germ line lineages (Figure 2). When used in conjunction with Essential 8 Medium, rhLaminin-521 enables a trouble-free transition from feeder-based media to a xeno-free, feeder-free media system.
Figure 1. Optimum recovery of PSCs during transition to Gibco Essential 8 Medium in the absence of small molecule inhibitors. An episomal CD34+-derived iPSC line was cultured on mouse irradiated feeders in KSR-based medium prior to transition to Essential 8 Medium. For transition, colonies were passaged using collagenase IV per the protocol provided with the Essential 8 Adaptation Kit and seeded into Essential 8 Medium in wells coated with rhLaminin-521 (green diamonds), Gibco Geltrex matrix (blue circles), or rhVTN-N (red triangles). Confluency of cultures was monitored using the IncuCyte ZOOM™ system post-passage (n=3 per condition).
Figure 2. Human ESCs maintain expression of self-renewal factors. Human embryonic stem cells (ESCs) were grown under feeder-free conditions using Gibco Essential 8 Medium for 20 passages in wells coated with rhLaminin-521. The cells were stained for pluripotency markers using the PSC 4-Marker ICC Kit.
Single-cell passaging with Essential 8
RevitaCell Supplement can be used for routine single-cell passaging of PSCs in Essential 8 Medium, providing efficient recovery of cells while retaining normal morphology, pluripotency, and karyotype. Furthermore, the prosurvival small molecule contained within this cocktail is shown to be a more selective ROCK inhibitor, minimizing the off-target pathways affected.
Figure 1. Gibco RevitaCell Supplement shows utility in single-cell passaging using Gibco TrypLE Select Enzyme. iPSCs generated using episomal vectors were passaged using TrypLE Select Enzyme and seeded at a density of 25,000 viable cells/cm2 onto culture plates coated with truncated recombinant human vitronectin, in Gibco Essential 8 Medium with 1X RevitaCell Supplement. Following a 24-hour recovery, iPSCs were fed with Essential 8 Medium alone for the remainder of their time in culture.
Figure 2. Normal morphology is retained following single-cell passaging using Gibco Essential 8 Medium plus Gibco RevitaCell Supplement. iPSCs generated using episomal vectors were passaged using Gibco TrypLE Select Enzyme and seeded at a density of 25,000 viable cells/cm2 onto culture plates coated with truncated recombinant human vitronectin, in Essential 8 Medium with 1X RevitaCell Supplement. Following a 24-hour recovery, iPSCs were fed with Essential 8 Medium alone for the remainder of their time in culture.
StemFlex
Superior clonal expansion following single-cell passaging
Figure 1. StemFlex Medium supports PSCs in recovery from flow sorting, demonstrating as much as 5-fold improvement in clonal expansion following single-cell passaging in the absence of ROCK inhibitor. PSCs expanded in StemFlex Medium on the rhLaminin-521 substrate for >3 passages were singularized using TrypLE Select reagent. They were then subjected to flow cytometric sorting to isolate live pluripotent stem cells (Tra1-60+/propidium iodide (PI)-negative) and were seeded at 1, 3, or 5 cells/well in a 96-well plate. Following plating, cells were fed with fresh medium every 3 days, and the percentage of wells achieving >5% confluency by day 14 was assessed via whole-well imaging on the IncuCyte ZOOM system. These data indicate that StemFlex Medium provides superior performance in the cells’ recovery from this stressful event.
Improved reprogramming workflow and robust colony formation
Figure 2. StemFlex Medium provides an improved workflow and robust colony formation for easy clone selection following reprogramming of somatic cells. Human dermal fibroblasts were reprogrammed using the Invitrogen CytoTune-iPS 2.0 Sendai Reprogramming Kit. Beginning on day 8 (upon introduction of the stem cell medium per the standard workflow), cells were fed either daily with mTeSR1 Medium or every other day using the StemFlex Medium system. (A) Robust colony formation was observed for the StemFlex Medium system. (B) Improved reprogramming efficiency was also observed for all three donors fed every other day using StemFlex Medium, relative to mTeSR1 cultures on a daily feed schedule.
Robust support of PSC cultures during expansion and maintenance
Figure 3. StemFlex Medium provides long-term maintenance of normal PSC properties. Culture displays normal morphology (left); a normal karyotype is observed after 20 passages on a weekend-free flexible feeding schedule (right).
Essential 8 Flex
Figure 1. FGF2 activity over time in PSC culture medium. Unlike other feeder-free PSC culture media, Essential 8 Flex Medium has been optimized to extend the activity of unstable components such as FGF2. Extended activity allows for routine culture without the daily feeding.
Figure 2. Long-term stability of cells cultured in Essential 8 Flex medium. Healthy karyotypes of pluripotent stem cells cultured in Essential 8 Flex medium have been observed after long term culture of cells.
Robust pluripotent stem cell cultures
FGF2 levels widely fluctuate in standard PSC culture medium, even with daily feeding (Figure 3). The use of Essential 8 Flex medium significantly limits these fluctuations, producing more robust, stable culture conditions with the added value of a flexible feeding schedule.
‡ This promotion is available only to customers in Austria, Belgium, Czech Republic, Denmark, Finland, France, Hungary, Italy, Germany, Luxembourg, Netherlands, Norway, Poland, Portugal, Slovakia, South Africa, Spain, Sweden, Switzerland, and United Kingdom who currently use human pluripotent stem cells in their research. Trial request discounts are at the discretion of the Technical Sales Specialist and must be received by March 31, 2020. Orders must include the relevant promotional code. All pricing information and values based on current list price Limit one trial discount per customer. The trial discount includes 80% discount for new customers and 50% discount for existing customers on their choice of stem cell media Essential 8 (A1517001) Essential 8 Flex (A2858501) or StemFlex (A3349401). Cannot be combined with other discounts or promotions. The discount can only be used once per customer. Void where prohibited by local law, company policy or customer policy. Other restrictions may apply. Please note that the value of the specific sample the customer may receive from the manufacturer under this program can be considered as a gift or an advantage, and as a result pursuant to applicable local laws or regulations, it could give rise to declaration, notification, publication or reporting (in particular, such advantage shall be published if the beneficiary is a French entity falling within the scope of the French “Loi Bertrand”). The customer also warrants it will comply with any local law or regulation and with any professional or ethical code or internal regulation relating to transparency or forbidding it to receive such free item, as the case may be.
For Research Use Only. Not for use in diagnostic procedures.